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Scallop MyGPx gene promoter cloning and expression

Author: DanZhongGuo
Tutor: HouLin；LiuWeiDong
School: Liaoning Normal University
Course: Marine biology
Keywords: Scallop Selenium dependent glutathione peroxidase mRNA expression Promoter analysis SNP
CLC: S917.4
Type: Master's thesis
Year: 2011
Downloads: 27
Quote: 0
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Abstract


Scallop (Mizuhopecten yessoensis) is an economically important shellfish, are widely distributed in Japan's northern island of the coast of the northern peninsula, Sakhalin and the four northern islands. In 1982, imported from Japan to China, owing to its individual large, and high market value, in Dalian and other regions will soon become an important shellfish species. The last decade, there has been growing scallop slow seedling production is low and mass death and other issues, so that scallop aquaculture has been a huge economic losses, has seriously hampered the development of China's shellfish industry. However, exploration of the immune system to shellfish, a brief introduction MyGPx gene helps us effectively control scallops disease, in order to better develop shellfish aquaculture. In this paper, gene cloning methods to scallop mantle cDNA as a template, by RT-PCR and RACE technology has been scallop glutathione peroxidase (MyGPx) full-length cDNA sequence (GenBank number: HQ433529 ). MyGPx gene is 1081bp, including 28bp of the 5 'untranslated region, 603bp open reading frame containing polyadenylation signal (AATAAA) of 450bp3' non-coding region. Multiple sequence alignment showed, MyGPx coding sequences and other invertebrates and vertebrates selenium dependent GPx has a very high affinity. Phylogenetic analysis shows scallop, pearl oyster bivalve Pu, mussel of GPx are associated with GPx1 and GPx2 has a high homology. By gene walking method to obtain 1686bp gene sequence that includes 58bp MyGPx cDNA sequence, 1628bp of the 5 'flanking region. Using TRANSFAC and AliBata 2.1 software predicted MyGPx 5 'flanking sequence contains transcription factor binding sites is assumed. By comparing the 15 scallops MyGPx 5 'flanking sequence and found nine SNP loci is assumed that these SNP loci discovery of molecular breeding scallop provides more potential molecular marker loci. By real-time quantitative PCR analysis MyGPx in different tissues and different stages of embryonic development and anguillarum (Vibrillo anguillarum) after stimulation with each time period relative expression levels. Studies have found a body in the mantle MyGPx highest expression during embryonic development in 10 trochophore larval period in the highest expression, bacteria stimulated 12 hours experimental group and the control group was the largest amount of difference. We can deduce, GPx may be an important growth factors and immune factors in the adult scallop tissue growth, embryonic development and larval immune defense plays a vital role.

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CLC: > Agricultural Sciences > Aquaculture, fisheries > Aquatic basic science > Aquatic Biology > Aquatic Zoology
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